Effect of Overnight Staining on the Quality of Flow Cytometric Sorted Stallion Sperm: Comparison with Tradtitional Protocols

Flow cytometry is considered the only reliable method for the separation of X and Y chromosome bearing spermatozoa in equines. The MoFlo SX DP sorter is highly efficient, allowing the production of foals of the desired sex. However, to achieve acceptable pregnancy rates the currently used protocol requires working with fresh semen obtained close to, or at, the sorting facility. An alternative protocol was tested during two consecutive breeding seasons. Fresh stallion semen was cooled for 20 h, during which staining with Hoechst 33342 took place. On the following day, this sample was flow sorted and compared with spermatozoa from the same ejaculate that had been sexed on the previous day. All sperm parameters evaluated remained unchanged when fresh sorted and refrigerated sorted semen were compared. Pre‐sorting storage at 5°C did not alter sperm velocities nor kinetics, viability or membrane permeability, production of reactive oxygen species, mitochondrial membrane potential or DNA fragmentation index of the sorted sample. The findings open for the possibility of using semen from stallions housed far from the sorting facilities. Processed and stained sperm could be shipped refrigerated on the previous day, sorted and inseminated on the next day.     

Giant cell tumor of the accessory carpal bone in a dog

A 4-year-old Springer Spaniel was referred because of a 6-week history of progressive left forelimb lameness. A marked pain response was elicited by palpation over the left accessory carpal bone. Radiography revealed a lytic, markedly expansile lesion of the accessory carpal bone. The bone was excised, and pancarpal arthrodesis was performed. Histologic examination of the excised bone revealed giant cell tumor. At 14 months after surgery, the dog was using the limb normally. Radiography of the carpus revealed satisfactory progression of the arthrodesis, and thoracic radiographs were negative for metastases.     

Improved Prediction of Ovulation Time may Increase Pregnancy Rates to Artificial Insemination in Lactating Dairy Cattle

A prospective observational study was conducted in two Australian dairy herds to assess the potential for improving pregnancy rates (proportions of inseminations that result in pregnancy) to artificial insemination (AI) if the time of ovulation could be predicted with more certainty. Herd 1 calved year‐round and inseminations were performed during two periods each day. Herd 2 calved during autumn–winter and inseminations were performed only after the morning milking each day. In both herds, the AI to ovulation interval of enrolled cows was determined by trans‐rectal ovarian ultrasonography approximately 0, 12, 24 and 36 h after AI, and pregnancy was assessed by palpation per rectum 35–56 days after AI. Also, in Herd 1 vaginal electrical resistance (VER) measurements were taken at approximately 0, 12, 24 and 36 h after AI, and in Herd 2 cows were fitted with neck mounted activity meters that monitored cow activity count in 2‐h periods. There was substantial variation in the intervals from AI to ovulation within and between herds (mean ± SD 21.2 ± 10.7, n = 102; 14.7 ± 10.4, n = 100 in herds 1 and 2, respectively). Pregnancy rates were higher for inseminations close to, but preceding, ovulation. Using combined herd data (n = 202), the highest pregnancy rate (50.8%) was observed for inseminations between 0 and 16 h before ovulation, a period in which only a modest proportion of inseminations (31.2%) occurred. In contrast, pregnancy rate was significantly lower (28.7%; risk ratio 0.6; 95% CI 0.4–1.0; p = 0.039) for inseminations between 16 and 32 h before ovulation, a period where the highest proportion of inseminations (53.2%) occurred. Thus pregnancy rates could potentially be improved if a greater proportion of inseminations were conducted shortly before ovulation. In Herd 1, mean VER during the peri‐ovulatory period varied with time from ovulation. Lowest values (mean ± SEM, VER = 64.8 ± 1.2, n = 55) occurred approximately 18 h before ovulation and were significantly lower than measurements approximately 6 h before ovulation (67.4 ± 1.0; n = 73; p = 0.003). Further work is required to determine if VER can be used to identify ovulation time and hence the optimal time to inseminate in individual animals. In Herd 2 a modest proportion of inseminations (26.9%) occurred between 24 and 40 h after the onset of increased cow activity where the highest pregnancy rate (67.9%) was observed, whereas a significantly lower pregnancy rate (42.4%; risk ratio 0.6; 95% CI 0.4–0.9; p = 0.036) was observed for inseminations between 8 and 24 h after the onset of increased cow activity where the highest proportion of inseminations (56.7%) occurred. Thus cow activity monitoring may be useful to identify the optimal time to inseminate cows. Results from this study indicate that improved methods of ovulation prediction may allow better insemination timing relative to ovulation and consequently increased pregnancy rates.     

The effects of induced parturition in dairy cows on the incidence of mortality in calves from commercial herds in south-western Victoria

SUMMARY The effects of induced parturition on calf mortality rates were studied in 24 winter-calving, pasture-fed, dairy herds in south-western Victoria. Parturition was induced when most cows were between 27 and 35 weeks of pregnancy. Control groups were selected from cows in the same herds in which calving was not induced. The incidence of mortality within 7 days of birth was much higher among calves from cows in induced groups relative to those in control groups (estimated median herd mortality incidence 72% and 7%, respectively). The incidence of calf mortality was high among groups induced at all stages of pregnancy that were investigated. However, differences in calf mortality incidence, relative to controls, were increased among groups induced at earlier stages of pregnancy.     

Mercury contamination of the harbour porpoise Phocoena phocoena and other cetaceans from the North Sea and the kattegat

Total Hg levels were determined in 17 harbour porpoises Phocoena phocoena collected in the North Sea and the Kattegat (muscle, liver and kidney), as well as in a sperm whale Physeter macrocephalus, a common dolphin Delphinus delphis and two bottle-nose dolphins Tursiops truncatus collected in Belgium. Juvenile porpoises (total body length less than 140 cm) were characterized by total Hg concentrations increasing with length (t.i. with age) with median values of 3.1 μg g^?1 dw in muscle, 6.2 in liver and 5.7 in kidney, and liver to muscle resp. liver to kidney ratios varying between 1 and 2. In adults, liver concentrations were higher (up to 500 μg g^?1 dw) with liver to muscle and to kidney ratios up to 20. Median McHg concentrations in 5 of the porpoises were 4.0 μg g^?1 dw in muscle and 6.9 in liver. These values did not increase as strongly with length (t.i. with age, at least for juveniles) as total Hg did; as a consequence, the relative McHg levels decreased from 100% in juveniles to 2 or 3 % in the liver of adults. Such results seem to reflect the existence of a slow mineralization phenomenon and storage of inorganic Hg, mainly in the liver.     

Redox cycling induces spermptosis and necrosis in stallion spermatozoa while the hydroxyl radical (OH•) only induces spermptosis

Oxidative stress is a major factor explaining sperm dysfunction of spermatozoa surviving freezing and thawing and is also considered a major inducer of a special form of apoptosis, visible after thawing, in cryopreserved spermatozoa. To obtain further insights into the link between oxidative stress and the induction of apoptotic changes, stallion spermatozoa were induced to oxidative stress through redox cycling after exposure to 2‐methyl‐1,4‐naphthoquinone (menadione), or hydroxyl radical formation after FeSO₄ exposure. Either exposure induced significant increases (p < 0.05) in two markers of lipid peroxidation: 8‐iso‐PGF_2α and 4‐hydroxynonenal (4‐HNE). While both treatments induced changes indicative of spermptosis (caspase‐3 activation and decreased mitochondrial membrane potential) (p < 0.01), menadione induced sperm necrosis and a dramatic reduction in motility and thiol content in stallion spermatozoa. Thus, we provided evidence that oxidative stress underlies spermptosis, and thiol content is a key factor for stallion sperm function.     

Serological status of cattle to bovine viral diarrhoea virus and bovine herpesvirus 1 at entry to and exit from Australian feedlot backgrounding facilities

A total of 6195 cattle were enrolled in this observational study. Serum antibody concentrations to bovine herpesvirus 1 (BHV1) and bovine viral diarrhoea virus (BVDV) were measured at entry to and exit from backgrounding facilities to assess their statuses on arrival and the extent of seroconversion to these viruses during backgrounding. The backgrounding facilities were contiguous with five feedlots in: Queensland (two sites), New South Wales, South Australia and Western Australia. Cattle were held in the backgrounding facilities for a minimum of 29 days and a median of 34 days. On backgrounding facility entry, 32.7% of the study population was seronegative to BVDV, but 85.7% was seronegative to BHV1. After commingling in the backgrounding facilities, of the cattle that were seronegative on backgrounding facility entry, 33.9% and 30.3% showed a serological increase to BVDV and BHV1, respectively. At backgrounding facility exit, when cattle were placed in their feedlots, 19.6% and 59.1% were seronegative to BVDV and BHV1, respectively, and 0.26% were persistently infected with BVDV. There was a strong association between seroincrease to BVDV and seroincrease to BHV1 (P = 0.005) at animal level in cohorts known to contain an animal persistently infected with BVDV.